dc.date.accessioned | 2013-03-12T12:38:58Z | |
dc.date.available | 2013-03-12T12:38:58Z | |
dc.date.issued | 2004 | en_US |
dc.date.submitted | 2004-10-19 | en_US |
dc.identifier.citation | Bratbak, Daniel Fossum. Gamma-glutamyl cysteine synthetase and L-buthionine-S,R-sulfoximine (BSO): a new selection strategy for gene-transduced neural and hematopoietic stem/progenitor cells.. Prosjektoppgave, University of Oslo, 2004 | en_US |
dc.identifier.uri | http://hdl.handle.net/10852/29473 | |
dc.description.abstract | In most experimental gene therapy protocols involving stem/progenitor cells, only a small fraction of cells, often therapeutically inadequate, can be transduced and express the therapeutic gene. A promising strategy for overcoming this problem is the use of a dominant selection marker, such as a drug-resistance gene. In this paper, we have explored the potential of the heavy subunit of gamma-glutamyl cysteine synthetase (gamma-GCSh) to act as a selection marker. We found that 3T3 fibroblasts transduced with a bicistronic retroviral vector, coding for gamma-GCSh and the enhanced green fluorescent protein (eGFP), were highly resistant to L-buthionine-S,R-sulfoximine (BSO), a gamma-GCS inhibitor with a very low clinical toxicity profile. The level of resistance was not proportional to the increase in intracellular glutathione. In fact, cells overexpressing both heavy and light gamma-GCS subunits had higher intracellular GSH levels, and a lower level of resistance to the cytotoxic activity of BSO compared to cells overexpressing gamma-GCSh alone. 3T3 fibroblasts overexpressing gamma-GCSh could be selected from cultures containing both naïve and gene-modified cells by application of exogenous BSO selection pressure for 4 days. Also primary neural stem/progenitor cells derived from the lateral ventricles of mouse neonatal brains and primary hematopoietic stem/progenitor cells from mouse bone marrow, transduced with the gamma-GCSh-eGFP vector, could be selected by BSO treatment in vitro. These results provide proof-of-principle that neural and hematopoietic stem/progenitor cells, transduced with a potentially curative gene and co-expressing the simultaneously transduced gamma-GCSh gene, can be selected by treatment with BSO. | nor |
dc.language.iso | eng | en_US |
dc.subject | medisinsk biokjemi | |
dc.title | Gamma-glutamyl cysteine synthetase and L-buthionine-S,R-sulfoximine (BSO): a new selection strategy for gene-transduced neural and hematopoietic stem/progenitor cells. | en_US |
dc.type | Master thesis | en_US |
dc.date.updated | 2004-11-23 | en_US |
dc.creator.author | Bratbak, Daniel Fossum | en_US |
dc.subject.nsi | VDP::726 | en_US |
dc.identifier.bibliographiccitation | info:ofi/fmt:kev:mtx:ctx&ctx_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&rft.au=Bratbak, Daniel Fossum&rft.title=Gamma-glutamyl cysteine synthetase and L-buthionine-S,R-sulfoximine (BSO): a new selection strategy for gene-transduced neural and hematopoietic stem/progenitor cells.&rft.inst=University of Oslo&rft.date=2004&rft.degree=Prosjektoppgave | en_US |
dc.identifier.urn | URN:NBN:no-36957 | |
dc.type.document | Prosjektoppgave | en_US |
dc.identifier.duo | 21458 | en_US |
dc.contributor.supervisor | Lorico, Aurelio | en_US |